In vivo analysis of endodermal and dorsal forerunner cell migration

内胚层和背侧先行细胞迁移的体内分析

• 批准号: 10552236
• 负责人: Stephanie Woo
• 金额: $ 11.86万
• 依托单位: UNIVERSITY OF CALIFORNIA, MERCED
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10552236
• 关键词: Address; Adhesions; Adopted; Behavior; Binding Proteins; Bioinformatics; Candidate Disease Gene; Cells; Characteristics; Clustered Regularly Interspaced Short Palindromic Repeats; Congenital Abnormality; Cytoskeletal Proteins; Data; Data Set; Development; Disease; Dorsal; Embryo; Endoderm; Endoderm Cell; Epithelial; Exhibits; Failure; Feeds; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Goals; Heterogeneity; Individual; Knock-in; Label; Life; Light; Logic; Maps; Measurement; Mesenchymal; Molecular; Morphogenesis; Movement; Organ; Phenotype; Population; Process; Proteins; Regulation; Reporter; Resolution; SOX17 gene; Sampling; Shapes; Signal Transduction; Specific qualifier value; Technology; Time; Tissues; Transgenes; Translating; Vesicle; Work; Zebrafish; base; cell behavior; cell growth regulation; cell motility; cell type; cohesion; differential expression; epithelial to mesenchymal transition; gastrulation; imaging approach; in vivo; in vivo imaging; interest; migration; parent grant; protein expression; reconstitution; single-cell RNA sequencing; transcription factor; transcriptome sequencing; transcriptomics

PROJECT SUMMARY In the developing embryos, the formation of organs and tissues arises from the collective actions of many individual cells which may change their shape, size, adhesion, and motility. Our long-term goal is to understand how these cellular behaviors are regulated in both space and time to correctly build each tissue and organ. In this project, we will leverage two closely related yet phenotypically distinct cell types, the endodermal and dorsal forerunner cells in zebrafish embryos, to better understand the regulation between migratory mesenchymal states and coherent epithelia. During early gastrulation, endodermal cells exhibit mesenchymal characteristics; they are highly migratory, spatially dispersed, and avoid contact with each other. In contrast the dorsal forerunner cells are tightly associated with each other and display epithelial characteristics, even as they migrate collectively. Remarkably, both cell types are specified by the same developmental signals and express many of the same marker genes, despite their divergent migration behaviors. In Aim 1 of this project, we will use single- cell RNA sequencing and bioinformatic analysis to compare the transcriptional profiles of endodermal and dorsal forerunner cells and identify differentially expressed candidate genes likely involved in the regulation of cell migration modes. In Aim 2, we will generate split fluorescent protein knock-in zebrafish lines to characterize the expression and localization of key endodermal and dorsal forerunner genes. This approach is based on technology developed in the parent grant to this diversity supplement application. If successful, this study will identify key genes responsible for different modes of migration in two closely related, yet distinct cell types. Our findings will shed light on the broader question of how small differences in transcriptional states translate into major differences in cellular behavior.

项目摘要 在发育中的胚胎中，器官和组织的形成源于许多人的集体作用 可能会改变其形状，大小，粘附和运动性的单个细胞。我们的长期目标是了解 这些细胞行为如何在空间和时间上调节，以正确构建每个组织和器官。在 这个项目，我们将利用两种密切相关但表型不同的细胞类型，内胚层和背侧 斑马鱼胚胎中的先驱细胞，以更好地了解迁移间充质之间的调节 国家和相干上皮。在早期胃肠道期间，内皮细胞表现出间充质特征。 它们是高度迁移的，在空间上分散，避免彼此接触。相比之下 细胞彼此紧密相关并显示上皮特征，即使它们迁移 集体。值得注意的是，两种细胞类型均由相同的发育信号指定，并表达许多 尽管迁移行为不同，但同一标记基因的基因。在该项目的AIM 1中，我们将使用单一 细胞RNA测序和生物信息学分析，以比较内胚层和背侧的转录特征 先驱细胞并鉴定可能与细胞调节有关的差异表达的候选基因 迁移模式。在AIM 2中，我们将生成分裂的荧光蛋白敲入斑马鱼线以表征 钥匙内皮和背前先驱基因的表达和定位。这种方法基于 父母在这种多样性补充应用中开发的技术。如果成功，这项研究将 确定在两种密切相关但不同的细胞类型中，负责不同迁移模式的关键基因。我们的 调查结果将阐明更广泛的问题，即转录状态的小差异如何转化为 细胞行为的主要差异。